摘要
以EMBL 3 DNA为载体,构建肝豆状核变性(HLD)患者的基因组文库。从HLD患者白细胞提取高分子量DNA,经Sau 3A部分酶解,电泳回收15～23kb DNA片段,EMBL 3 DNA用BamHI及EcoRI进行双酶解,两者经T_4 DNA连接酶连接成重组DNA分子。从E.coli溶原菌BHB2688及BHB2690用冻融法及超声破碎法提取包装蛋白,包装效率为1～8x10^7pfu/μg EMBL 3 DNA。连接分子用包装蛋白体外包装,获得了9.2×10~6个重组子,达到建库要求。
Hepatolentioular degeneration (HLD) is an autosomal recessive disorder of copper metabolism characterized by copper accumulation in different tissues. The HLD locns is mapped at 13ql4 and linked to several DNA markers. We used EMBL 3 DNA bo construct its genomio DNA library. Purified EMBL 3 DNA was digested by Bam HI and Eco El, and ligated to Sau 3A digested HLD DNA fragments to form recombinant DNA. The ligation reaction was in vitro packaged by packaging extracts, which were prepared by the freeze/thaw and sonioation method from E.coli lysogens BHB2688 and BHB2690. 9.2×105 recombinant phagos were obtained. The genomic library may be used to search for RFLP probes and the disease gene.
出处
《上海医科大学学报》
CSCD
1990年第5期331-334,共4页
Journal of Fudan University(Medical Science)
关键词
肝豆状核变性
重组DNA
基因组文库
Hepatolentieular degeneration
EMBL 3 DNA
DNA ligation
in vitro package
geaomie library
